The primary goal of this bridging grant is to establish a comprehensive register of the inflammatory responses elicited in macrophages by microbial stimuli. We intend to dissect the responses to elucidate the underlying regulatory mechanisms controlling inflammatory responses in macrophages and to identify key regulators and effectors that may be useful pharmaceutical targets. Currently funded projects in the lab focus specifically on the mechanisms by which Toll-like receptors (TLRs) recognize ligands and stimulate pro-inflammatory signals. These existing projects (like many in the field) will benefit greatly from the establishment of"Glue" core resources (ie the reagents core), but these projects are necessarily limited by their focus on the mechanisms of actions of individual proteins; we do not presently have the all technical expertise and resources to draw larger maps of macrophage inflammatory responses that would allow us, and others in the Glue consortium, to identify novel proteins that might be crucial for developing a complete understanding of the innate immune response in macrophages. Working with the genomics, proteomics, and bioinformatics cores, we will fully characterize the genes and proteins whose expression are actively regulated during macrophage detection of bacterial products that stimulate the cells through individual TLRs and combinations of TLRs. Specifically we will examine the responses to LPS (TLR4), peptidoglycan (TLR2/TLR6), bacterial DNA (TLR9) and bacterial flagellin (TLR5), as well as whole Gram-negative and Gram-positive bacteria and yeast. Using perturbational analysis of pro-inflammatory signaling networks, we will analyze the relationships between clusters of genes that are coordinately regulated, and in the process we expect to identify novel components that, by virtue of their association with known key elements of the response, might be critical to the pathologies associated with inflammation.